Yuka Sasaki, Yuna Mogi, Mizuki Yoshioka, Ke Liu, Yuichi Otsuka
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引用次数: 0
摘要
毒素-抗毒素(TA)基因模块控制着细菌的各种活动。具有不同功能的新型毒素仍在不断被发现。本研究旨在确定肠出血性大肠杆菌 O157 编码的 ECs3274-ECs3275 基因对是否具有 TA 系统的功能。为了描述这个假定的 TA 系统,我们分析了表达 ECs3274、ECs3275 或两者的大肠杆菌的生长情况;使用细菌腺苷酸环化酶双杂交试验分析了 ECs3274 和 ECs3275 之间的相互作用;以及使用凝胶移动性转移试验分析了 ECs3274 的 DNA 结合能力。我们观察到,ECs3274 抗毒素与 ECs3275 毒素相互作用,被 Lon 蛋白酶破坏稳定性,并通过其螺旋-转向-螺旋(HTH)基团抑制其启动子活性。这些特性与典型的 II 型 TA 抗毒素一致。有趣的是,ECs3275具有其他TA毒素所没有的HTH基序,并且是ECs3275毒性所必需的,这表明ECs3275可能通过调节特定基因的表达来发挥其毒性。
A type II toxin-antitoxin system, ECs3274-ECs3275, in enterohemorrhagic Escherichia coli O157.
The toxin-antitoxin (TA) genetic module controls various bacterial events. Novel toxins with different functions are still being discovered. This study aimed to determine whether the ECs3274-ECs3275 gene pair encoded by enterohemorrhagic Escherichia coli O157 functions as a TA system. To characterize this putative TA system, we analyzed the growth of E. coli expressing ECs3274, ECs3275, or both; the interaction between ECs3274 and ECs3275 using bacterial adenylate cyclase two-hybrid assays; and the DNA-binding ability of ECs3274 using gel-mobility shift assays. We observed that the ECs3274 antitoxin interacted with the ECs3275 toxin, was destabilized by Lon protease, and repressed its promoter activity via its helix-turn-helix (HTH) motif. These properties are consistent with those of typical type II TA antitoxins. Interestingly, ECs3275 has an HTH motif not observed in other TA toxins and is necessary for ECs3275 toxicity, suggesting that ECs3275 may exert its toxicity by regulating the expression of specific genes.
期刊介绍:
Bioscience, Biotechnology, and Biochemistry publishes high-quality papers providing chemical and biological analyses of vital phenomena exhibited by animals, plants, and microorganisms, the chemical structures and functions of their products, and related matters. The Journal plays a major role in communicating to a global audience outstanding basic and applied research in all fields subsumed by the Japan Society for Bioscience, Biotechnology, and Agrochemistry (JSBBA).